Dengue viruses (category A pathogen) are estimated to infect 100 million people annually and are considered to cause one of the most important arthropod-borne viral diseases in terms of human morbidity and mortality. Although 40% of the world's population can benefit from a dengue vaccine, there is no clear pathway to commercialization of such a vaccine. Barriers to commitment of the necessary funds to drive commercialization derive from scientific, market, medical and investment-funding uncertainties that, added together, create unacceptable risk for today's companies. Dengue and dengue hemorrhagic fever are caused by one of four closely related virus serotypes. Infection with one of the serotypes does not provide cross-protective immunity from the other serotypes and is believed to exacerbate the disease upon infection with a second serotype. To date, no dengue vaccine has been licensed and human clinical trials of several dengue vaccine candidates have not demonstrated durable immunity against all four dengue serotypes. Hawaii Biotech, Inc. has developed a multicomponent, recombinant subunit dengue vaccine that has the potential to elicit long-lasting immunity to all four dengue serotypes. The vaccine will be developed for phase I clinical trials to be performed in collaboration with The Walter Reed Army Institute of Research. The vast majority of the preclinical development work outlined in this proposal will be performed under Good Manufacturing Practice (cGMP) and Good Laboratory Practice (GLP). The preclinical development work encompasses scale-up and cGMP manufacture of recombinant dengue antigens covering the four dengue serotypes, purification of the antigens, testing and release of the vaccine prior to use in humans, toxicology testing of the vaccine in animal models, fill/finish of the vaccine active pharmaceutical ingredient (API) and adjuvant API and preparation of an Investigational New Drug Application.